Absorbent article

ABSTRACT

An absorbent article has an absorbent material and an outer surface including an inhibitor which is partially bound to the absorbent material and substantially inhibits the colonization of bacteria within the absorbent article. 
     An absorbent article has an absorbent material. The absorbent material has an outer surface. The absorbent article includes a pre-toxin limiting agent which is partially bound to the absorbent material and substantially retards the production of bacteria-produced toxins by the bacteria residing within the absorbent article. 
     An absorbent article has an absorbent material. The absorbent material has an outer surface. The absorbent article includes a toxin enclosing agent. The toxin enclosing agent substantially inhibits the migration of toxin outwardly from within the absorbent article towards the outer surface of the absorbent article.

CROSS REFERENCE TO RELATED APPLICATION

This application is a continuation application of U.S. application Ser.No. 11/725,462, filed Mar. 19, 2007, which is a continuation applicationof U.S. application Ser. No. 10/744,361, filed Dec. 22, 2003, which is acontinuation of International Application No. PCT/US02/20617, filed Jun.28, 2002, designating the U.S., which claims the benefit of U.S.Provisional Application No. 60/302,507, filed Jun. 29, 2001.

FIELD OF INVENTION

The present invention relates to a tampon which employs mechanisms toreduce the potential risk of Toxic Shock Syndrome (“TSS”) associatedwith tampon wear or any other absorptive product such as surgical gauzeor any product whose use has been directly associated with toxic shocksyndrome, such as but not excluded to diaphragms and cervical cups.

BACKGROUND OF THE INVENTION

Disposable absorbent devices for the absorption of human exudates arewidely used. These disposable devices typically have a compressed massof absorbent formed in the desired shape, which is typically dictated bythe intended consumer use. For example, in the case of menstrualtampons, the device is intended to be inserted at least partially into abody cavity for absorption of the body fluids generally dischargedduring a woman's menstrual period.

There exists in the human female body a complex process which maintainsthe vagina and physiologically related areas in a healthy state. Infemales between the age of menarche and menopause, the normal vaginaprovides an ecosystem for a variety of microorganisms, which must bemaintained in a relatively delicate balance. Bacteria are thepredominate type of microorganisms present in the vagina, and most womenharbor about 10⁹ bacteria per gram of vaginal secretions. The bacterialflora of the vagina is comprised of both aerobic and anaerobic bacteria.The more commonly isolated bacteria include Lactobacillus species,Corynebacteria species, Gardnerella vaginalis, Staphylococcus species,Peptococcus species, aerobic and anaerobic Streptococcal species, andBacteroides/Preuotella species. Other microorganisms that have beenisolated from the vagina on occasion include yeast (Candida albicans),protozoa (Trichomonas vaginalis), mycoplasma (Mycoplasma hominis),chlamydia (Chlamydia trachomatis), and viruses (Herpes simplex). Theselatter organisms are generally associated with vaginitis or sexuallytransmitted diseases, although they may be present in low numberswithout causing symptoms.

Physiological, social and idiosyncratic factors affect the quantity andspecies of bacteria present in the vagina. Physiological factors includeage, days of the menstrual cycle, and pregnancy. For example, vaginalflora present in the vagina throughout the menstrual cycle can includeLactobacilli, corynebacterium, ureaplasma, and mycoplasma. Social andidiosyncratic factors include presence and method of birth control,sexual practices, systemic disease (e.g., diabetes), and medication.

Bacterial proteins and metabolic products produced in the vagina canaffect other microorganisms and the human host. For example, generallythe pH of the vagina between menstrual periods is mildly acidic, havinga pH ranging from about 3.8 to about 4.5. This pH range is generallyconsidered the most favorable condition for the maintenance of normalflora. At that pH, the vagina normally harbors the numerous species ofmicroorganisms in a balanced ecology, playing a beneficial role inproviding protection and resistance to infection and making the vaginainhospitable to some species of bacteria such as S. aureus. The low pHis a consequence of the growth of Lactobacilli and their production ofacidic products. Microorganisms in the vagina can also produceantimicrobial compounds such as hydrogen peroxide and bacteriocins whichattack and eliminate other bacterial species. One example is thelactocins, bacteriocin-like products of Lactobacilli directed againstother species of Lactobacilli. Some microbial products may affect thehuman host. For example, S. aureus can produce and excrete into itsenvironment a variety of exoproteins including enterotoxins, toxic shocksyndrome toxin-1 (“TSST-1”), and enzymes such as protease and lipase.

Vaginal menstrual toxic shock syndrome is a rare syndrome characterizedby rapid onset of high fever, vomiting, diarrhea, and rash followed by arapid drop in blood pressure and vital organ failure. TSS is associatedwith the presence of S. aureus bacteria and one or more exotoxins whichare produced by the bacteria. The exotoxins associated with TSS includebut may not be limited to Streptococcus: Exotoxin A, Exotoxin B,Exotoxin C and Staphylococca: Pyrogenic Exotoxin C, Enterotoxin A,Enterotoxin B, Enterotoxin C, Enterotoxin F, and TSST-1. Usingtraditional culture based techniques, S. aureus has been identified inthe vagina of approximately 16% of healthy women of menstrual age(Recent clinical studies using DNA based techniques have shown thisnumber to be much higher). It has been found that approximately 10% ofthe S. aureus isolated from the vagina are capable of producing TSST-1.TSS is not caused by the bacteria per se but rather by the toxic effectsof the associated exotoxin which can pass from the vagina and otherinternal body cavities into the blood stream.

TSS has been associated with the use of absorbent pads within the vaginawhich may promote the growth of bacteria and the production of exotoxinin their vicinity. The syndrome has been observed with surgicaldressings, and is also associated with the use of catamenial tampons.The syndrome appears to occur with elevated frequency in associationwith those absorbent pads which are characterized by high levels ofabsorbency and which accordingly are left inside the body for extendedperiods.

While a preferred approach for reducing the risk of TSS when usingabsorbent pads is proper use and frequent changes of new pads for usedones, various other approaches have been proposed by the art forreducing the risk of TSS associated with an internal absorbent pad. Oneapproach is the incorporation of antimicrobial or bacteriocidal agentsinto the absorbent pad such as the use of iodine bactericides in tamponsand catamenial sponges. Such an approach is not always suitable for usein the catamenial product, however, because a bactericide which isactive against S. aureus can adversely affect other beneficial bacteriawhich make up the vaginal flora, thereby upsetting the healthy balancediscussed above. Another related method describes the use of catamenialtampons comprising substances such as organic acids which will maintaina pH of about 4.5 to about 2.5 in the fluids absorbed during the use ofthe tampon such that the growth of pathogenic bacteria is inhibited.

Other approaches are directed to inactivation of the TSS toxin such asthe administration of L-ascorbic acid for the detoxification of the S.aureus toxins, Pyrogenic, Exotoxin C (Schlievert) and StaphylococcalEnterotoxin F (Bergdoll) TSS-1. While this method does not ascribe amechanism for the effectiveness of ascorbic acid at neutralizing TSS-1,it observes that L-ascorbic acid is known to be a reducing agent andstrong antioxidant and that it may operate to inactivate bacterialtoxins by reducing disulfide bonds within the toxins.

Another approach is directed to the incorporation of substances withinan absorbent pad which inhibit the production of TSS exotoxins by S.aureus. This method describes the incorporation of non-toxic divalentmagnesium cations in absorbent pads to reduce the concentrations ofavailable magnesium binding ions below those critical for optimalproduction of TSST-1 and other staphylococcus products.

Despite these developments, there remains a desire in the art forabsorbent pads suitable for internal use, including catamenial tampons,which are characterized by improved immobilization of TSST-1 toxingenerated within the absorbent product without adversely affecting thenormal vaginal flora.

SUMMARY OF THE INVENTION

The present invention encompasses an absorbent article having anabsorbent material and an outer surface including an inhibitor which isat least partially bound to the absorbent material and substantiallyinhibits the colonization of bacteria within the absorbent article.

The present invention can also encompass an absorbent article having anabsorbent material. The absorbent material has an outer surface. Theabsorbent article includes a pre-toxin limiting agent which is at leastpartially bound to the absorbent material and that substantially retardsthe production of bacteria-produced toxins by the bacteria residingwithin the absorbent article.

In another embodiment, an absorbent article has an absorbent material.The absorbent material has an outer surface. The absorbent articleincludes a pre-toxin limiting agent which is at least partially bound tothe absorbent material and a pre-toxin limiting agent that substantiallyde-activates bacteria produced toxins arising from bacteria residingwithin the absorbent article.

In yet another embodiment, an absorbent article has an absorbentmaterial. The absorbent material has an outer surface. The absorbentarticle includes a toxin enclosing agent. The toxin enclosing agentsubstantially inhibits the migration of toxin outwardly from within theabsorbent article towards the outer surface of the absorbent article.

In another aspect, an absorbent article has an absorbent material. Theabsorbent material has an outer surface. The absorbent article includesa temperature limiting agent. The temperature limiting agentsubstantially inhibits the elevation of temperatures within theabsorbent article.

In yet another aspect, an absorbent article has an absorbent material.The absorbent material has an outer surface. The absorbent articleincludes a temperature equilibrium agent. The temperature equilibriumagent substantially reduces the rate at which the absorbent articleraises above equilibrium in an ambient environment.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates the mechanism for ATP production within a cell.

FIG. 2A illustrates gas level as a function of time for non-menstrualconditions.

FIG. 2B illustrates gas level as a function of time for menstrualconditions.

FIG. 3A illustrates toxin level as a function of temperature.

FIG. 3B illustrates temperature as a function of time for a tampon.

FIG. 4 illustrates the production of the octapeptide, its release intothe environment and its signaling for the production of the toxin.

DETAILED DESCRIPTION OF THE INVENTION

The physiological/microbial activity occurring in the tampon duringmenstrual wear indicates a possibility that if the toxin producingstrain of S. aureus is present it may produce toxin within or near thesurface of the tampon. Moreover, the toxin initiates disease by cominginto contact with and penetrating the vaginal mucosa. The risk of TSScan be reduced by 1) preventing S. aureus from colonizing the tampon; 2)altering the conditions within the tampon to insure that the perceivedrequired conditions for toxin production are not present; 3) disruptingthe epithelial or antigen presenting cell and T-cell binding sites forthe toxin; 4) and/or retaining the toxin within the tampon.

Section A will provide terms which will assist the reader in bestunderstanding the features of the invention and not to introducelimitations in the terms not consistent with the context in which theyare used in this specification. Theses definitions are not intended tobe limiting.

There are three phases to prevent a diseased state. Section B willdiscuss Phase I which involves the decrease in and/or prevention ofsurface or internal bacterial colonization of the tampon. Section C willdiscuss Phase II which addresses potential toxin production by theTSST-1 producing strain of S. aureus and therefore attempt to inhibitproduction of toxin within and/or on the surface of the tampon. SectionD will discuss Phase III in which the goal is to deactivate the bindingsite of the toxin and/or prevent the toxin from coming into contact withthe vaginal mucosa when the tampon is colonized by the TSST-1 producingstrain of S. aureus and it is exposed to the required conditions fortoxin production.

A. Terms

In general in this specification, the term “tampon” is used to refer toa finished tampon typically after a compression process and to any typeof absorbent structure that is inserted into the vaginal canal or otherbody cavities for the absorption of fluid therefrom.

As used herein the terms “vaginal cavity,” “within the vagina,” and“vaginal interior,” are intended to be synonymous and refer to theinternal genitalia of the human female in the pudendal region of thebody. The term “vaginal cavity” as used herein is intended to refer tothe space located between the introitus of the vagina (sometimesreferred to as the sphincter of the vagina) and the cervix and is notintended to include the interlabial space, including the floor ofvestibule. The externally visible genitalia generally is not includedwithin the term “vaginal cavity” as used herein.

As used herein, the term “bound” means less than about 10% of thebiological activity associated with inhibiting toxin production orgrowth of S. aureus in the tampon is lost by soaking the tampon over an8 hour period at 100° F. in three times the syngnya capacity of sterilephysiologic saline solution. The saline is removed from the tampon bycompressing the tampon at 1 psi on a series of blotter paper until ≦0.5grams of fluid is absorbed by the blotter paper on the lastcompression/absorption sequence and remeasuring the biological activityremaining in the tampon. The extraction and testing should be conductedin the absence of light. The ratio of initial biological activity of thetampon pre-extraction to post extraction is >90%.

As used herein, the term “partially bound” means less than about 50% ofthe biological activity associated with inhibiting toxin production orgrowth of S. aureus in the tampon is lost by soaking the tampon over an8 hour period at 100° F. in three times the syngnya capacity of sterilephysiologic saline solution. The saline is removed from the tampon bycompressing the tampon at 1 psi on a series of blotter paper until ≦0.5grams of fluid is absorbed by the blotter paper on the lastcompression/absorption sequence and remeasuring the biological activityremaining in the tampon. The extraction and testing should be conductedin the absence of light. The ratio of initial biological activity of thetampon pre-extraction to post extraction is >50%.

As used herein, the term “substantially bound” means less than about 25%of the biological activity associated with the toxin production organicgrowth associated with S. aureus of the tampon is lost by soaking thetampon over an 8 hour period at 100° F. in three times the syngnyacapacity of sterile physiologic saline solution. The saline is removedfrom the tampon by compressing the tampon at 1 psi on a series ofblotter paper until ≦0.5 grams of fluid is absorbed by the blotter paperon the last compression/absorption sequence and remeasuring thebiological activity remaining in the tampon. The extraction and testingshould be conducted in the absence of light. The ratio of initialbiological extraction to post extraction is >75%.

As used herein, the term “biological activity” of a tampon is measuredby the method described by J. Parsonnet, et. Al. (J. Infect. Dis. 1996,173:98-103).

As used herein, the term “encapsulation” means the surrounding off or“caging” of a compound using a physical or chemical component.

As used herein, the term “nonabsorbent” means a non-absorbing componentof an absorbent article, as distinct from the absorbing article itself,and that individual component (particle or fiber) will not swell orabsorb fluid. Typically, this means less than a 0.15% weight gain of thecomponent when placed in a saline solution for 10 minutes and thenremoved and compressed at 1 psi repeatedly against a series of blotterpapers until less than about 0.5 gram of saline is absorbed by theblotter paper on the last compression/absorption sequence. As usedherein an absorbent material, as distinct from an absorbent component isa material from which the absorbent core is made and may hold fluid inthe intercapillary fibers or voids and may itself be an absorbent ornon-absorbent component.

As used herein, the term “inhibitor” means any agent that prevents thenormal growth of an organism or the activity of an enzyme or protein.

As used herein, the term “pre-toxin limiting agent” means any agent thatprevents the initiation or actual production of a toxin.

As used herein, the term “deactivation” means to make less toxic ornontoxic.

As used herein, the term “permeable structure” means any article thatselectively allows components of a particular size less than or equal tothe particular size to readily pass through without substantialresistance. An overwrap is an example of a permeable structure.

B. Phase I: Reduce and/or Prevent Colonization of the Tampon by VaginalMicroorganisms

To reduce the risk of TSS, antimicrobial agents are incorporated withinan absorptive product such as a tampon to prevent the colonization ofthe tampon by bacterial species. If the bacterial species colonize, thebacteria can create toxin. If S. aureus is present and the toxin iscreated, the woman is at risk for TSS. The tampon represents a newecological niche within the vaginal environment for the indigenousbacterial species of the vagina to colonize. New understandings of thisindigenous population point toward the hypothesis that the greatestdiversity, especially of minor species, in the vaginal biofilm/colonyoccurs at the surface of the vaginal mucosa. The highest probability ofS. aureus initial colonization occurs at this vaginal mucosasurface/tampon interface. Components secreted by other species ofvaginal bacteria can amplify toxin production by S. aureus. Therefore,to reduce the risk of an increase in population of S. aureus, via growthin the tampon, and extracellular components secreted by other organismsfrom affecting the potential rate of toxin produced, the presentinvention incorporates inhibitors that inhibit colonization into thetampon's design and construction. Section i. discusses the incorporationof antimicrobial agents into the absorptive product. Section ii.discusses the incorporation of antifouling agents into the absorptiveproduct. Section iii. discusses the incorporation of biostatic agentsinto the absorptive product. Section iv. discusses the incorporation ofnegatively charged molecules. Finally, Section v. discusses theencapsulation of antimicrobial agents and biostatic agents.

i. Antimicrobial Agents

Antimicrobial agents may be incorporated into the construction of theabsorptive, non-absorptive, and/or overwrap components of the tampon.Antimicrobial agents are agents which inhibit growth or kill bacteria.Controlling the localization of these antimicrobial agents is important.If the localization of the antimicrobial agents is not controlled,disruption and/or depletion of the indigenous microflora covering thevaginal mucosa would potentially be harmful to the overall health of thevaginal canal. The antimicrobial activities of many antimicrobial agentsare dependent upon solubilization and uptake by the microorganisms ofinterest.

The antimicrobial agents may be covalently linked to the non-absorbentmaterial of the tampon, either directly or through modification. Thepresence and detection of the antimicrobial agents by the vaginalmicroorganisms would be sufficient to delay or decrease the potentialoverall colonization of the tampon during typical wear. Modification ofthe antimicrobial agent to allow the antimicrobial to be immobilized,for example via —CH₂— or —CH₂—CH₂— etc. linker, while not obstructingtheir active sites would allow for increased availability andeffectiveness against colonization of the tampon. However, linking theanitmicrobial agents to the absorbent component of the tampon maydecrease the tampon's overall effectiveness.

Antimicrobial agents that can be utilized for this purpose may includebut are not limited to Clindamycin, Lysostaphin, PHMB, Triclosan, andquaternary ammonia compounds. Clindamycin and Lysostaphin are discussedin more detail below.

An additional benefit of using Clindamycin is that this agent depressestoxin production by S. aureus. Clindamycin hydrochloride, or CLEOCIN®HCl, is currently used to treat vaginal infections. It is asemisynthetic antibiotic produced by 7(S)-chloro-substitution of the7(R)-hydroxyl group of the parent compound lincomycin. Clindamycininhibits bacterial protein synthesis at the level of the bacterialribosome. It binds preferentially to the 50S ribosomal subunit andaffects the process of peptide chain initiation. Although clindamycinphosphate is inactive in vitro, rapid in vivo hydrolysis converts thiscompound to the antibacterially active form. Clindamycin has been shownto be an effective antimicrobial agent for a number of organismsreported to be associated with bacterial vaginosis, such as Bacteroidesspp., Prevotella spp., Gardnerella vaginalis, Mycoplasma hominis andPeptostreptococcus spp.

The antimicrobial agent lysostaphin (a zinc dependent, 25-kDaglycyl-glycine endopeptidase isolated from S. simulans) is a specificpeptidoglycan hydrolase (“PGH”) enzyme for Staphylococcus bacteria. PGHenzymes are incorporated and coordinated with the use of absorbentarticles to reduce the risk of colonization of the absorbent article byStaphylococcus. PGH enzymes are naturally present in small amounts inbacteria where they are essential for cell wall re-modeling that mustoccur with cell growth and division. Because bacterial peptidoglycanstructures differ significantly, PGHs can be found to target specificbacterial species or groups of related organisms. Specifically, theaction of lysostaphin is hydrolysis of the —Gly+Gly— bond in thepentaglycine inter-peptide link joining Staphylococcal cell wallpeptidoglycans, thereby, digesting the peptidoglycan “structuralbarrier” of the bacterial cell walls. Added in excess they rapidlydigest peptidoglycan resulting in cell rupture. Therefore, lysostaphin,which is a specific PGH enzyme to Staphylococcal organisms, wouldprevent S. aureus and other Staphylococcal organisms from colonizing thetampon without adversely affecting the non-Staphylococcal dominatespecies of the vaginal canal. Lysostaphin's N-terminal of the enzymaticprotein is the enzymatically active region and the C-terminal is whatconfers target cell specificity and allows it to distinguish between S.aureus and its parent cells S. simulans (Baba & Schneewind, EMBO J.1996). Crosslinking the protein to a matrix via its C-terminal regionwould anchor the protein and still allow it to some degree to beenzymatically active against Staphylococcal spp.

ii. Antifouling Agents

Antibiofilm formation agents may be incorporated into an absorbentproduct such as a tampon to prevent the colonization of the absorbentproduct. Antifouling agents are an antibiofilm formation agent which mayalso be incorporated into the tampon's construction. The antifoulingagents can be incorporated into the tampon's construction by binding.The binding of the antifouling agents is important because of the nonimpact or influence upon an environment external to its currentenvironment. For example, an anti-microbial agent located in the tamponshould not impact or influence the growth of bacteria outside thetampon.

The two types of antifouling agents which may be incorporated areFuranones and L-acyl homoserine lactones. Furanones are halogenatedcompounds that are known to depress gram-negative bacterial growth andkill gram-positive bacteria. L-acyl homoserine lactones are specific togram-negative bacteria and are also known to control virulence factors.These agents need to be immobilized to the components of the tampon andprohibited from coming into direct contact with the microflora of thevaginal canal. Furthermore, quorum sensing signals are active biofilmforming signals for gram-positive organisms and may be employed toinhibit colonization of a tampon. Quorum signals will be discussed indepth in Section C of the Phase II discussions.

iii. Biostatic Agents

Biostatic agents decrease bacterial growth. Biostatic agents such as thecommon histochemical dyes, methylene blue, and gentian violet can becross-linked to cellulose fibers and other potential absorptivematerials. Methylene blue is a redox dye that raises the oxygenconsumption of cells. This means that the protons (hydrogens) of thematerials to be oxidized are passed on to the oxygen molecules presentin the environment. Methylene blue in an unbound state acts as anelectron carrier short circuiting the electron transport process whichis responsible for ATP (adenosine triphosphate) production within thecell (as shown in FIG. 1). A similar mechanism is hypothesized whenmethlene blue is bound to a fiber structure.

iv. Negatively Charged Molecules

Coating the absorbent article and its components with negatively chargedmolecules prevents the colonization of the absorbent product/tampon.Negatively charged tampon and/or its components electrostatically repelbacterial cell surfaces which are negatively charged. If the materialsof the absorbent product were negatively charged via the use of PO₄ orSO₄ etc., bacteria would be repelled from forming biofilm and or rapidlydividing in this negatively charged environment.

v. Encapsulation of Antimicrobial Agents and Biostatic Agents

Incorporating the encapsulation of enzymes and or active agents with anabsorptive product such as a tampon prevents the colonization of S.aureus. Antimicrobial and/or biostatic agents can be encapsulated by theuse of hydrophilic isocyanate polymers. These polymers are water-solubleand a variety of molecules can be incorporated into these hydrophilicurethane type polymers. During the polymerization of hydrophilicisocyanate prepolymers, carbon dioxide is typically generated keepingthe oxygen levels in the immediate environment low. This low oxygenenvironment aides in the overall stability of any enzyme added to theprepolymer emulsion which becomes an integral part of the resultantcopolymer. The enzyme(s) or other chemical antimicrobial/biostaticagent(s) participates in the polymerization and is typically chemicallyattached at multiple points within the resulting isocyanate copolymermatrix essentially becoming caged within the isocyanate polymer. Asdescribed previously, the overall availability and effectiveness of thecaged enzyme may be increased by first modifying the attachment site ofthe enzyme to the polymer matrix via the addition of a “linker”molecule. Within this copolymer matrix a variety of enzymes have beenshown to be thermally stable over time. Since this is an aqueouspolymerization event, upon hydration of the copolymer matrix the enzymeor microbial agent becomes “active” and can exhibit a retention ofspecific activity as high as 80% of “uncaged” activity. Thissolubilization of the isocyante polymer allows for a controlled releaseof the enzyme as fluid triggers the solubilization of the urethane likegel. The components, which exhibit antimicrobial or biostatic activity,are covalently bound to the polymer. Upon hydration, the polymer iseroded to release the active material at the preferred site of action.

It is preferred to keep antimicrobial and biostatic agents isolated fromthe vaginal mucosa. This may be accomplished by the use of appropriateoverwraps that cover the absorbent core. For example, formed filmoverwraps may be used or nonwoven overwraps made of rayon, cotton,polyesters, polyolefins that provide separation where the core isisolated from the surrounding vaginal tissue preventing destructionand/or alteration to the vaginal flora. The overwrap may also act incombination with the absorbent core to provide a one-way valve such thatfluid enters the tampon and is irreversibly trapped. This tamponexhibits the one-way property when it is loaded with saline at 75% ofsyngyna capacity, allowed to equilibrate for fifteen minutes and thenplaced on blotter paper, the tampon is then rolled over the blotterpaper without pressure applied to remove free fluid on the surface ofthe tampon. The tampon is then placed on a fresh blotter paper andcompressed with a force of 1 psi. If the squeeze out on the blotterpaper is less than 0.1 grams the tampon is considered to be a one-wayvalve.

Physical separation of the inhibitor or antimicrobial agent can bedemonstrated by adding sterile saline solution (at syngyna capacity)gently blot the surface to remove free fluid and place the tamponcontaining the inhibitor onto an agar place colonized with a lawn ofLactobacillas. If the Lactobacillas lawn is not disturbed/killed at thecontact point between the tampon and the agar plate physical separationis demonstrated.

C. Phase II: Prevent Toxin from being Produced by TSST-1 Producing

Extensive in vitro work describes the environmental and geneticconditions required for S. aureus to produce TSST-1 toxin. Current invivo physiological understanding of the dynamics of dissolved oxygen andcarbon dioxide in the tampon during menstrual wear indicate that optimumconditions for toxin production by S. aureus must exist within thetampon. The oxygen required for potential toxin production within thetampon is believed to come from the inherent air within the tampon andthe oxygen carried by the menstrual fluid absorbed by the tampon. Theinherent oxygen of the tampon is essentially unutilized by thefacultative microorganisms of the vaginal canal in the absence ofblood/menses. However, in the presence of a heavy loading ofblood/menses virtually all of the oxygen in the tampon can be consumed,while the dissolved carbon dioxide levels within the tampon risesignificantly above that present in the vaginal environment.

As shown in FIG. 2A, during non-menstrual wear, carbon dioxide levels ofthe tampon rose to be essentially equivalent to those observed in thevagina. However, as shown in FIG. 2A, the oxygen levels within anon-menstrual tampon remained essentially at atmospheric levels.

As shown in FIG. 2B, during menstruation, the mean levels of carbondioxide in about 50% of the tampons exceed the carbon dioxide levels ofthe vaginal environment. The mean levels of oxygen within the tampondecrease with apparent loading of the menses such that about 50% of themenstrual tampons reached an oxygen level near the ranges of oxygenlevels observed in the vagina after 8 hours of wear. The decline ofoxygen and the increase of carbon dioxide within a menstrual tampon maybe at least partially related to microbial metabolic activity. Sincemenses is a mixture of venous and arterial blood, some of the oxygen maybe “lost” due to partitioning from the gas phase to the liquid phasewhere it binds to un-oxygenated heme molecules present in the menses.The oxygen and carbon dioxide levels observed in a menstrual tamponafter prolonged wear are consistent with in vitro data showing highlevels of toxin production by S. aureus under similar O₂/CO₂ conditions.

Phase II provides several approaches to prevent toxin from beingproduced by the TSST-1 producing strains of S. aureus. Particularly,pre-toxin limiting agents are used to retard the production of bacteriaproduced toxins. Part i. discusses oxygen scavengers incorporated intothe construction of the tampon. Part ii. discusses the alteration of theheme in menses. Part iii. discusses the encapsulation of antimicrobialagents via hydrophilic isocyanate polymers. Part iv. discusses theconstruction and/or additive to an absorbent structure, which eithermoderates (i.e. reduces) the thermal energy released during expansion ofa tampon and/or cools the absorbent article. Part v. discusses theblockage of either the production of the octapeptide (Quorum Signal) orthe binding of the peptide to the cell membrane receptor.

i. Oxygen Scavengers

In a number of in vitro experiments, oxygen has been shown to beessential for toxin production by S. aureus. An oxygen scavengerincorporated into the construction of a tampon would significantlyreduce, if not entirely prohibit, toxin production within the product.Examples of oxygen scavengers, which could be employed, are antioxidantssuch as ascorbic acid, tocopherol and retinal. Compounds such as theantioxidants used in food substances such as butylated hydroxyanisole,di-tertiary-butyl-patacresol, propyl gallate, phenylenethiourea, andaldoalpha-napthylamine could also be used. Other oxygen scavengingagents employed may be composed of transition metal complexes, chelatesof a salicyclic acid, salicylate salt, metal complex, and/or chelate ofan organic polycarboxylic acid preferably an amino polycarboxylic acidwherein the transition metal could be supplied via the iron present inmenses from the degraded heme molecules present in this body fluid. Thistype of oxygen scavenging molecule is typically activated by contactwith water or water vapor.

These same oxygen-scavenging agents have the additional benefit ofmoderating the pH of the tampon. Because optimum toxin production occursat a neutral pH, any agent which will lower the pH of the internalenvironment of the tampon into the acidic range could effectively reducetoxin production. Such agents are organic acids, for example ascorbicacid, polycarboxylic acid, etc. Not only does ascorbic acid act as ameans to lower the pH and/or scavenge oxygen, but the ascorbic acid alsodeactivates the toxin. While the mechanism of deactivation is unknown,the present application includes the use of ascorbic acid in a tampon todeactivate the potential presence of TSST-1 toxin to the point ofrendering it non-toxic, i.e. not lethal in animal experiments.

ii. Alteration of the Heme in Menses

Another approach is to alter the heme in menses such that it has a loweraffinity for oxygen. This may be accomplished by placing BPG(2,3-bisphosphoglycerate) within the tampon. BPG lowers the affinity ofoxygen for hemoglobin by a factor of 26. This is the compound used todissociate oxygen from the heme enabling the unloading of oxygen totissue capillaries in the body. If the oxygen can not associate withheme, it is probable that it will no longer be bioavailable to themicroorganisms potentially colonizing the tampon such as S. aureus andtherefore, avoid the perceived required conditions for toxin productionwithin the tampon during menstruation.

iii. Encapsulation of Antimicrobial Agents via Hydrophilic IsocyanatePolymers

In Phase I, there was a discussion of encapsulation of antimicrobialagents via hydrophilic isocyanate polymers to prevent colonization ofthe tampon. This same technology can be utilized to encapsulate agentswhich would prevent or decease the production of toxin by S. aureus. Analternative method to the isocyanate polymer encapsulation would be theuse of alginates precipitated from solution. Material that may bephysically encapsulated includes but is not limited to carbon black,antimicrobials, oxygen scavengers (i.e. ascorbic acid), quorum signalanalogs and/or blockers, methylene blue, chitosan malate, etc. Thesesame compounds can be added to absorbent foams either pre- orpost-polymerization. Other agents which can be added include but are notlimited to ascorbic acid, tocopherol, glycerol, etc., all of which havebeen shown to reduce toxin production and/ or deactiviate the toxinTSST-1. Algenic acid can also be made into fibers via reaction withcellulosic fibers and provides a matrix for the additional cross-linkingof a number of compounds such as chitosan malate, which is known todepress microbial growth of S. aureus and significantly reduce itsability to produce the TSST-1 toxin. Thus, alginate particles/fibers actas an active agent carrier that can be used with or in a component ofthe construction of an absorbent article such as a tampon to depressmicrobial growth of S. aureus and significantly reduce its ability toproduce the TSST-1.

iv. Construction and/or Additive to an Absorbent Structure which eitherModerates or Reduces the Thermal Energy Released during Expansion of aTampon and/or Cools the Absorbent Article

A potential risk factor for toxin production in vivo is believed to beelevated temperature. In an attempt to moderate or depress any tendencythe tampon may have to exist at an elevated temperature in vivo, anumber of compounds could be used. As shown in FIG. 3A, in in vitroexperiments a one degree Fahrenheit increase in temperature can resultin a 50% increase in TSST-1 production by S. aureus.

This patent application claims any technique of construction and/oradditive to an absorbent structure, which either moderates (i.e.reduces) the thermal energy released during expansion of a tampon and/orcools the absorbent article (either of compressed or non-compressedconstruction) such that it preferably remains at or below typical bodytemperature of 98.6° F. If the internal tampon temperature does riseabove the ambient body temperature of 98.6° F., it returns to 98.6° F.within 30 minutes.

A number of currently marketed products are tested for their potentialchange in internal temperatures upon hydration, i.e. a measure of theirabsorbent material's heat of dissolution. The experimental designinvolves the use of non-lubricated condoms submerged with the open endabove water into a 98.6° F. water bath. A thermocouple is placed in asmall hole drilled through the middle of a tampon, which is insertedinto the condom such that it too is submerged into the water bath. Asecond thermocouple is placed between the tampon and the surroundingcondom. The condom-surrounded tampon is then allowed to come to thetemperature of 98.6° F. Sterile saline is also allowed to come to atemperature of 98.6° F. in the same water bath. A gush of the 98.6° F.sterile saline (at syngyna capacity) is then pumped in such that thetampon becomes saturated. The internal and external tampon temperatureis monitored. In addition to commercially available products, a tamponconstructed of a super absorbent material as well as one constructed offoam absorbent material (FAM) is tested.

As shown in FIG. 3B, no changes were observed for the tampons “external”temperature.

As shown in FIG. 3B, currently marketed products that are constructed asa compressed “plug” of absorbent material exhibited a sharp rise ininternal temperature upon hydration with fresh menses. The thermalenergy is a result of the release of expansion energy upon hydration ofthe compressed absorbent plug structure. Referring to FIG. 3B, note thatthe temperature of the commercially available products quickly returnedto water bath temperature after the action of hydration. The tamponcomposed of super absorbent material exhibited an internal temperatureof over 2° F. above that of the water bath and sustained thistemperature for a prolonged period of time of 7 hours. Temperatureincrease of the super absorbent tampon may be related to the heat ofdissolution of the materials used in its construction, a phenomenapreviously not considered in tampon construction. The foam absorbentmaterial (“FAM”) constructed tampon's internal temperature rose onlyslightly and quickly returned to the original ambient temperature of98.6F.

The Epson Salt or magnesium sulfate hepta-hydrate (MgSO₄—H₂O) issomewhat endothermic in nature and could be employed to counteract anyphysical reactions to elevate temperature within the tampon. Epsom Saltor magnesium sulfate hepta-hydrate (MgSO₄—H₂O) has a low energy ofdissolution. Certain absorbent materials such as psyllium husk andcertain FAMs have been shown to moderate the temperature of a tamponunder the experimental conditions outlined above such that thetemperature of the tampon is not significantly elevated (as shown inFIG. 3A and FIG. 3B). Other agents that possess zero or less than zeroheats of dissolution include but are not limited to NaCl, Ca(NO₃)₂—4H₂O,Na₂CO₃—10H₂O, CaCl₂—6 H₂O and various other magnesium salts.

An absorbent foam such as FAM or another absorbent may be saturated witha solution of salt such as CaCl2, MgCl2, or sodium ascorbate or absorbicacid and then dried leaving a residual salt or acid content within thefoam. It has been found that concentrations of these materials on aweight percent of foam between 0.1% and 10% can inhibit TSS-1production.

The architecture of the tampon being either a compressed plug or a loosesack has been demonstrated to impact the internal temperature of thetampon upon hydration. The loose sack design moderates the temperatureof the tampon such that its internal environment does not exceed thetemperature of the body during absorption of bodily fluids. In thisdesign cellulose could also be employed in the construction of theproduct to act to establish a less dense product to allow for bettertemperature control, i.e. body temperature or less.

v. Blockage of either the Production of the Octapeptide (Quorim Signal)or the Binding of the Peptide to the Cell Membrane Receptor

This patent application claims the use of such signals and/or signalanalogs/antagonist, either natural or artificial in origin to repressthe signal for the ultimate production of TSST-1 by S. aureus in anabsorbent article and or menstrual cup, diaphragm or other like devices.Likewise any agent that can either prevent the production of theoctapeptide, bind up the octapeptide after its production such that itis no longer available to “feed back” to the cell or occupy the bindingsites on the cell membrane such that the cyclic peptide—signalingmolecule octapeptide (agr D/autinducing peptide) can not bind and signalfor toxin production in the absorbent articles.

The gene cluster or operon that regulates the expressions of the toxindesignated TSST-1 is an example of Quorim Sensing. When an infectiousagent such as S. aureus invades a host, it first attaches and attemptsto grow and form a biofilm. The agr system controls the production ofmany secreted proteins: positively regulating enterotoxins,epidermolytic toxins and enzymes produced by staphylococci, andnegatively controls such proteins such as Protein A, fibronectin-bindingprotein and coagulase. During this time signals are sent out into theimmediate environment where the biofilm is forming. When enough of thissignal is present, it binds to a membrane receptor on the S. aureuscells to indicate that a critical density of bacteria has been reachedand cells of S. aureus may be released into the environment to continuethe process of infection. Therefore, when the population of the cells isdense enough and the concentration of signal is high enough such that anindividual bacterial cell can detect the signal toxin synthesis isinitiated. This high level of signal is referred to, as a “Quorum” ofsignal, the signal compound in the case of TSST-1 is an octapeptide autoinducing peptide. Toxin production occurs due to a cascade ofphosphorylation events in response to the quorum signal within the cellthat brings about the activation of the promoter region, which controlsthe expression of the genes encoded for the toxin. After the biofilm“burst”, the S. aureus cells adhere to a new site within the host andthe “Quorum Sensing” process begins again. FIG. 4 depicts the productionof the octapeptide, its release into the environment and its signalingfor the production of the toxin. (Arvidson, S. European Conference onToxic Shock, September 1997)

As shown in FIG. 4, blockage of either the production of the octapeptideauto inducing peptide (Quorum Signal) or the binding of the peptide tothe cell membrane receptor, could essentially lock the door to toxinproduction. With the use of natural repressors for TSST-1 productionsuch as competing analogs or antagonists peptides. Therefore, thispatent application claims the application of such an analog orantagonist peptide/compound to the construction of and/or as an additiveto an absorbent article such as a tampon to reduce the expression ofTSST-1 toxin by S. aureus during the absorbent articles use.

D. Phase III: Disrupt Toxin Binding Sites and/or Prevent Toxin fromContacting Vaginal Mucosa

Phase III of this patent application claims any technique (i.e.application of technique to an absorbent article) or tampon constructionthat acts to retain/adsorb the toxin into/on the tampon preventing thetoxin and/or toxin producing strains of S. aureus from coming intocontact with the vaginal mucosa. To prevent any TSST-1 toxin productionfrom contacting the vaginal mucosa, the tampon will be designed tospecifically attract and bind TSST-1 toxin from and within the tamponfrom the immediate environment around the tampon or from blood/mensesentering the tampon.

Several ways have been developed to retain the toxin in the tampon by avariety of ligand binding sites. The binding of the specific toxins ontothe fiber surfaces of the tampon prevents them from contacting vaginalmucosal tissue, and thereby, is expected to provide some degree ofprotection to the wearer against the toxin. The toxin can be retained bya direct affinity to its own antibody or antibody fragment or a chemicalhapten resembling its own binding site. Specific polypeptides orfragments of polypeptides which show an affinity for the toxin can alsobe employed to directly link the toxin. Toxins may also adhere tospecific oligonucleotide sequences which create direct or combinatoriallibraries and screen them for binding activity to the toxin that canidentify these oligonucleotides. Indirectly the toxin can be linked bysaturating the environment with protein A.

There are a variety of methods to attach ligands to fibers and/ormaterials, such as: 1) solid matrix support though cullulosic fibers: 2)activation coupling chemistry with the use of sodium periodate: 3) sitedirected antibody coupling though carbohydrates: 4) coupling of antibodyfragments with sulfhydral residues: and 5) amplification of antibodybinding sites such as avidin coupled matrixes or biotinylated antibodiesor fragments.

The above-described ligands need not only be linked to the tampon'sabsorptive fibers/particles and/or overwrap and/or materials (i.e.psyllium, FAM, etc.). The toxin binding ligands may also be attached tonon-absorptive/or absorptive glass beads, zeolites and charcoal asexamples. In addition, the ligands may be attached to the inner surfaceof the tampon overwrap composed of formed film topsheet, 3-D films, highloft structures, low capillary force gradients (hollow fibers, etc.).These materials act not only as one-way flow valves, allowing flow intothe tampon but not out, but also as the final “net” to catch the toxinbefore coming into contact with the vaginal mucosa.

In order for the toxin to remain in the tampon once it becomes linked toa particular ligand, the ligand must be anchored to the particularabsorbent or non-absorbent material composing the tampon. If thebiologically active molecules is not bound or partially bound to theinternal matrix of the tampon there may be a risk of the toxin cominginto contact with the vaginal mucosa. Here again, the one-way flow valveoverwrap structure can be incorporated to the final “safety net” tocatch any toxin or toxin complex from coming into contact with thevaginal mucosa.

Overwraps that isolate or trap fluid in the tampon by providing aphysical separation between the fluid in the core and the top surface ofthe tampon or vaginal membrane are defined as “one way valves.”

All documents cited in the Detailed Description of the Invention are, inrelevant part, incorporated herein by reference; the citation of anydocument is not construed as an admission that it is prior art withrespect to the present invention

While particular embodiments of the present invention have beenillustrated and described, it would be obvious to those skilled in theart that various other changes and modifications can be made withoutdeparting from the spirit and scope of the invention. It is thereforeintended to cover in the appended claims all such changes andmodifications that are within the scope of this invention.

1. A vaginal device comprising an overwrap, said overwrap comprising aninhibitor which is at least partially bound to said overwrap andsubstantially inhibits the growth of bacteria within said device.
 2. Thevaginal device of claim 1, wherein said inhibitor is an antimicrobialagent.
 3. The vaginal device of claim 1, wherein said inhibitor is anantifouling agent.
 4. The vaginal device of claim 3, wherein saidantifouling agent is a quorum sensing compound.
 5. The vaginal device ofclaim 4, wherein said quorum sensing compound is a peptide.
 6. Thevaginal device of claim 4, wherein said quorum sensing compound is anoctapeptide.
 7. The vaginal device of claim 4, wherein said quorumsensing compound is a cyclic peptide.
 8. The vaginal device of claim 1,wherein said overwrap further comprises an antifouling agent.
 9. Thevaginal device of claim 1, wherein said overwrap has an inner surface,wherein said inhibitor is at least partially bound to said inner surfaceof said overwrap.
 10. The vaginal device of claim 1, wherein saidoverwrap further comprises a biostatic agent.
 11. The vaginal device ofclaim 10, wherein said biostatic agent is methylene blue.
 12. Thevaginal device of claim 10, wherein said biostatic agent is ahistochemical dye.
 13. The vaginal device of claim 10, wherein saidbiostatic agent is gentian violet.
 14. The vaginal device of claim 1,wherein said vaginal device is a tampon.
 15. A vaginal device comprisingan overwrap, said overwrap comprising a biostatic agent which is atleast partially bound to said overwrap and substantially inhibits thegrowth of bacteria within said device.
 16. The vaginal device of claim15, wherein said biostatic agent is methylene blue.
 17. The vaginaldevice of claim 15, wherein said biostatic agent is a histochemical dye.18. The vaginal device of claim 15, wherein said biostatic agent isgentian violet.
 19. The vaginal device of claim 15, wherein saidoverwrap has an inner surface, wherein said biostatic agent is at leastpartially bound to said inner surface of said overwrap.
 20. The vaginaldevice of claim 15, wherein said vaginal device is a tampon.